黄夏白冬消癌汤对Lewis肺癌小鼠Bax/Bcl-2/Caspase-3信号通路的影响

Effect of Huangxiabaidong Xiaoai decoction on Bax/bcl-2/Caspase-3 signal pathway in Lewis Lung cancer mice

目的观察黄夏白冬消癌汤对Lewis肺癌小鼠肿瘤生长及Bax/Bcl-2/Caspase-3信号通路的影响。方法选取40只雄性C57BL/6小鼠,于小鼠右侧腋窝皮下注射Lewis肺癌细胞悬液建立肺癌模型。将造模成功的Lewis肺癌小鼠随机分为4组,每组10只。模型组给予生理盐水灌胃(1次/d),顺铂组给予顺铂2 mg/kg腹腔注射(每2 d 1次),黄夏白冬消癌汤低、高剂量组分别给予2 g/mL和4 g/mL黄夏白冬消癌汤灌胃(1次/d),均连续干预14 d。测量各组干预2,4,6,8,10,12,14 d后肿瘤长径与短径,计算肿瘤体积;干预结束后脱颈椎处死小鼠,称瘤重,计算抑瘤率;取各组小鼠瘤组织,采用Western blot法检测瘤组织中NF-κBp65、p-NF-κB p65、Bax、Bcl-2、Caspase-3、Cleved-Caspase-3蛋白表达情况,采用qPCR法检测瘤组织中NF-κB p65、Bax、Bcl-2、Caspase-3 mRNA相对表达量。结果黄夏白冬消癌汤低剂量组干预6~14 d期间各时间点肿瘤体积及干预结束后瘤重与模型组比较差异均无统计学意义(P均>0.05);顺铂组及黄夏白冬消癌汤高剂量组肿瘤生长延缓,各时间点肿瘤体积及干预结束后瘤重均明显低于模型组(P均<0.05),抑瘤率均明显高于黄夏白冬消癌汤低剂量组(P均<0.05),顺铂组与黄夏白冬消癌汤高剂量组各时间点肿瘤体积、干预结束后瘤重、抑瘤率比较差异均无统计学意义(P均>0.05)。顺铂组及黄夏白冬消癌汤低、高剂量组p-NF-κB p65/NF-κB p65及NF-κB p65、Bcl-2 mRNA相对表达量均明显低于模型组(P均<0.05),Cleved-Caspase-3/Caspase-3均明显高于模型组(P均<0.05);顺铂组及黄夏白冬消癌汤高剂量组Bax/Bcl-2及Bax、Caspase-3 mRNA相对表达量均明显高于模型组(P均<0.05),黄夏白冬消癌汤低剂量组与模型组比较差异均无统计学意义(P均>0.05);黄夏白冬消癌汤低、高剂量组p-NF-κB p65/NF-κB p65及Bcl-2 mRNA相对表达量均明显高于顺铂组(P均<0.05),Cleved-Caspase-3/Caspase-3及Bax、Caspase-3 mRNA相对表达量均明显低于顺铂组(P均<0.05),NF-κB p65 mRNA相对表达量与顺铂组比较差异均无统计学意义(P均>0.05);黄夏白冬消癌汤高剂量组Bax/Bcl-2与顺铂组比较差异无统计学意义(P>0.05),黄夏白冬消癌汤低剂量组明显低于顺铂组(P<0.05)。结论黄夏白冬消癌汤可以通过上调Bax、Caspase-3和下调Bcl-2、NF-κB p65、p-NF-κB p65的表达以促进肺癌细胞凋亡,发挥抗肿瘤作用。

Objective It is to explore the influence of Huangxiabaidong Xiaoai decoction on tumor growth and Bax/Bcl-2/caspase-3 signaling pathway in Lewis lung cancer mice.Methods Forty male C57BL/6 mice were selected and subcutaneously injected with Lewis lung cancer cell suspension into their right armpit to establish lung cancer models.The successfully modeled mice with Lewis lung cancer were randomly divided into 4 groups,with 10 mice in each group.The model group was given normal saline by gavage(once a day),the cisplatin group was given cisplatin 2 mg/kg by intraperitoneal injection(once every 2 days),and the low-dose and high-dose groups of Huangxiabaidong Xiaoai decoction were respectively given 2 g/mL and 4 g/mL Huangxiabaidong Xiaoai decoction by gavage,all the groups were continuously intervened for 14 days.The long and short tumor diameters were measured after 2,4,6,8,10,12,14 days of intervention in each group,and the tumor volume was calculated.After the intervention,the mice were sacrificed by removing cervical vertebra,the tumor weight was weighed,and the tumor inhibition rate was calculated.The tumor tissues of the mice in each group were taken to detect the protein expression of NF-κBp65,p-NF-κB p65,Bax,Bcl-2,Caspase-3,Cleved-Caspase-3 in tumor tissues by Western blot method,and to detect the mRNA expression of NF-κBp65,Bax,Bcl-2,Caspase-3 by qPCR method.Results There was no significant difference in tumor volume at each time point during the 6~14 days of intervention and tumor weight after intervention between the low-dose Huangxiabaidong Xiaoai decoction group and the model group(all P>0.05),the tumor growth of the cisplatin group and the high-dose Huangxiabaidong Xiaoai decoction group was delayed,the tumor volume at each time point and tumor weight after the intervention were significantly lower than those of the model group(all P<0.05),and the tumor inhibition rate was significantly higher than that of the low-dose Huangxiabaidong Xiaoai decoction group(P<0.05),there was no significant difference in tumor volume at each time point,tumor weight after the intervention and tumor inhibition rate between the cisplatin group and the high-dose Huangxiabaidong Xiaoai decoction group(all P>0.05).The mRNA relative expression of p-NF-κB p65/NF-κB p65,NF-κB p65 and Bcl-2 in cisplatin group and low-dose and high-dose groups of Huangxiabaidong Xiaoai decoction were significantly lower than those in the model group(all P<0.05),while that of Cleved-Caspase-3/Caspase-3 was significantly higher than that in the model group(P<0.05);the mRNA relative expression of Bax/Bcl-2,Bax,Caspase-3 in the cisplatin group and the high-dose Huangxiabaidong Xiaoai decoction group were significantly higher than those in the model group(all P<0.05),and there was no significant difference between the low-dose Huangxiabaidong Xiaoai decoction group and the model group(P>0.05);the mRNA relative expression of p-NF-κB p65/NF-κB p65 and Bcl-2 in the low and high dose groups of Huangxiabaidong Xiaoai decoction were significantly higher than those of cisplatin group(all P<0.05),the mRNA relative expression of Cleved-Caspase-3/Caspase-3,Bax,Caspase-3 were significantly lower than those of cisplatin group(all P<0.05),there was no significant difference in the relative expression of NF-κB p65 compared with cisplatin group(P>0.05);there was no significant difference in Bax/Bcl-2 between the high-dose Huangxiabaidong Xiaoai decoction group and cisplatin group(P>0.05),but that in the low-dose Huangxiabaidong Xiaoai decoction group was significantly lower than that of cisplatin group(P<0.05).Conclusion Huangxiabaidong Xiaoai decoction can promote apoptosis of lung cancer cells,thus to treat for the cancer though up-regulating the expression of Bax and Caspase-3 and down-regulating the expression of Bcl-2,NF-κB p65 and p-NF-κB p65.